<ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> הרדימי עכברים עם isoflurane (4% לזירוז, 1.5-2% לניתוח) באמצעות נהלים שאושרו IACUC. חשוב זנב ו / או צובט את הבוהן משמשים על מנת להבטיח את החיה הוא מסומם לגמרי. </li><li style=";text-align:right;direction:rtl"> שימוש גוזם מכרסם, לגלח את השיער מהחלק האחורי של הצוואר עד העיניים. </li><li style=";text-align:right;direction:rtl"> מניחים את העכבר בתוך מסגרת stereotaxic, על שמיכה מים מחדש במחזור הניתוח. לאבטח היטב את הראש עם ברים האוזן. </li><li style=";text-align:right;direction:rtl"> החל משחה העין, על מנת למנוע עין של החיה מהתייבשות. </li><li style=";text-align:right;direction:rtl"> ניהול, תת עורי, dexamethasone (0.2 מ&quot;ג / ק&quot;ג) ו Carprofen (5 מ&quot;ג / ק&quot;ג) כדי למנוע התנפחות של המוח ו / או תגובה דלקתית, בהתאמה. </li><li style=";text-align:right;direction:rtl"> לפני תחילת הניתוח, לחטא את אזור ההפעלה ידי ניגוב העור עם שלושה swipes לסירוגין של אלכוהול 70% ו בבטאדין. </li><li style=";text-align:right;direction:rtl"> כל המכשירים כירורגית כבר מראש לעקר באמצעות מעקר חרוז זכוכית. באמצעות מספריים שהיו מעוקרות עם אתנול, להסיר את העור מעל הגולגולת, החל לחתוך אופקי לאורך כל הבסיס של הראש, ואחריו שני חתכים לכיוון מקורי, כמעט והגיע העפעפיים, ואז שני חתכים אלכסוניים כי להתכנס בבית קו האמצע. </li><li style=";text-align:right;direction:rtl"> ירידה של פתרון + אפינפרין לידוקאין מוחל בשלב זה על periosteum כדי למנוע דימום מוגזם או כאב. עם אזמל, לסגת periosteum את הקצוות של הגולגולת. כמו כן, קל לחזור על שרירי הגב של הצוואר. </li><li style=";text-align:right;direction:rtl"> לגרד בעדינות את האזור כולו חשוף של הגולגולת עם אזמל כדי ליצור משטח יבש. זה חשוב מאוד, שכן הוא יאפשר הדבק לדבוק טוב יותר כאשר מיושם מאוחר יותר. </li><li style=";text-align:right;direction:rtl"> ברגע אתר הדמיה נבחרה, אחד הוא מוכן ליצור את החלון גולגולתי. ראשית, בעדינות &quot;למשוך&quot; מעגל של כ - 4 מ&quot;מ קוטר עם המקדחה פנאומטי שיניים. </li><li style=";text-align:right;direction:rtl"> לאחר קידוח קלה, להחיל לידוקאין + פתרון אפינפרין שוב על פני השטח את הגולגולת. הפסיקו את הקידוח כאשר שכבה דקה מאוד של העצם נשאר. על ידי לחיצה בעדינות על מרכז craniotomy כדי להרגיש איך זה נותן דרך, אפשר בדרך כלל יודעים כי בשלב זה הוא הגיע. </li><li style=";text-align:right;direction:rtl"> תחת ירידה של יתרון מליחים לקיחת trabeculae העצם - המבנה הספוגי של העצם - להרים משם את craniotomy מהגולגולת עם מלקחיים קצה דק מאוד. מלח חשוב, כמו זה יעזור להרים את הגולגולת למנוע דימום של הדורה. </li><li style=";text-align:right;direction:rtl"> החל Gelfoam כי כבר ספוג בעבר מלוחים מאטר הדורה לעצור דימום קטן מדי פעם מתרחשת כאשר הגולגולת מוסר. </li><li style=";text-align:right;direction:rtl"> לאחר ייבוש השטח מאטר הדורה ולהבטיח כי יש דימום לא, בעדינות להניח כוס סטרילית 5 מ&quot;מ לכסות להחליק על גבי מאטר הדורה. (הערה: קבוצות אחרות גם מקום ירידה של נקודת התכה נמוכה agarose (1.2%) מעל הדורה ולשים את coverslip על גבי אגר). </li><li style=";text-align:right;direction:rtl"> החל טיפת דבק cyanocrylate מבוסס בחצי הכדור ההפוך על הגולגולת. בעזרת מחט, בעדינות להחיל את הדבק מסביב לחלון תוך הקפדה לא לשים אותו תחת זכוכית. דבק כעת ניתן להחיל בשכבה דקה על פני השטח הכולל של הגולגולת. </li><li style=";text-align:right;direction:rtl"> לאחר הדבק התייבש, לערבב אקרילי שיניים ולהחיל אותו בכל השטח הגולגולת, המכסה גם שפה קטנה של להחליק את המכסה, כדי לאבטח אותו. </li><li style=";text-align:right;direction:rtl"> לאחר הבטחת כיסוי להחליק, לבצע גם קטן סביב החלון עם אקריליק שיניים. כמו כן, להטביע בר טיטניום אקריליק שיניים. בר זה יהיה מאוחר יותר לשמש כדי לצרף את העכבר בבטחה לשלב של המיקרוסקופ הדמיה. חשוב להבטיח כי הבר ברמה, כך שיהיה מקביל עם החלון גולגולתי. הנחת פיסת נייר מתחת לבר יכולים להרשות הבר להישאר ברמה תוך אקרילי מתקשה. </li><li style=";text-align:right;direction:rtl"> אקריליק שיניים מותר לרפא (להקשות) למשך 10 דקות, ועד אז הבר טיטניום קבוע במקום. מניחים את חיה בכלוב חמים עד שהוא מתאושש. </li><li style=";text-align:right;direction:rtl"> לאחר ההתאוששות מן ההרדמה, החיה ניתן הדמיה באותו יום. </li></ol>

<ol>
	<li>Svoboda, K., Denk, W., Kleinfeld, D., Tank, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=+In+vivo+dendritic+calcium+dynamics+in+neocortical+pyramidal+neurons."> In vivo dendritic calcium dynamics in neocortical pyramidal neurons.</a> Nature. 85, 161-165 (1997).</li><li>Lendvai, B., Stern, E. A., Chen, B., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experience-dependent+plasticity+of+dendritic+spines+in+the+developing+rat+barrel+cortex+in+vivo.">Experience-dependent plasticity of dendritic spines in the developing rat barrel cortex in vivo.</a> Nature. 404, 876-881 (2000).</li><li>Trachtenberg, J. T., Chen, B., Knott, G. W., Feng, G., Sanes, J. R., Welker, E., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+in+vivo+imaging+of+experience-dependent+synaptic+plasticity+in+adult+cortex.">Long-term in vivo imaging of experience-dependent synaptic plasticity in adult cortex.</a> Nature. 420, 788-794 (2002).</li><li>Portera-Cailliau, C., Weimer, R. M., DePaola, V., Caroni, P., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diverse+modes+of+axon+elaboration+in+the+developing+neocortex.">Diverse modes of axon elaboration in the developing neocortex.</a> PLoS Biol. 3, (2005).</li><li>Holtmaat, A., Wilbrecht, L., Knott, G. W., Welker, E., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experience-dependent+and+cell-type-specific+spine+growth+in+the+neocortex.">Experience-dependent and cell-type-specific spine growth in the neocortex.</a> Nature. 441, 979-983 (2006).</li></ol>

כפי שהראינו את הוידאו וגם את הדמויות משלים, הכנת חלון גולגולתי, בשילוב עם שימוש במיקרוסקופ שני הפוטונים, הוא כלי רב עוצמה מאוד ללמוד in vivo את המבנה והתפקוד של הניאוקורטקס. הטכניקה דורשת הכשרה קפדני להכיר את האנטומיה הרלוונטית ואת ניתוחים בסדר ומיומנויות כי זה דורש הכנה. רק ניתוחי...

ברים הם טיטניום מחוייט

a craniotomy surgery procedure for chronic brain imaging

טכניקות ההדמיה נעשים יותר ויותר חשוב בתפקוד המוח במחקר. ביניהם, שני הפוטונים מיקרוסקופ סורק לייזר התפתחה שיטה שימושית מאוד, משום שהיא מאפשרת את המחקר של המוח שלמים לחיות. עם ההכנות המתאימות, טכניקה זו מאפשרת תצפית באזור קליפת המוח באותו כרוניים, מ דקות עד חודשים. בסרטון הזה, אנו מראים הכנה כרונית בתחום ההדמיה vivo של המוח באמצעות שני פוטונים במיקרוסקופ. טכניקה זו היה בתחילה חלוץ ידי דר &#39;קארל סוובודה, כיום הווארד יוז רפואי חוקר במכון Janelia משק חקלאי. תכשירים כמו זה המוצג כאן יכול לשמש הדמיה של המבנה neocortical (למשל, הדינמיקה הדנדריטים axonal), להקליט הפעילות העצבית באמצעות סידן רגיש צבעים, אל קליפת המוח תמונה הדינמיקה זרימת הדם, או מהותי לימודי הדמיה אופטית. הדמיה של דיפ הניאוקורטקס אפשרי עם אופטימלית ניתוחים חלון גולגולתי. הפעלה בתנאים סטריליים ביותר האפשרית כדי למנוע זיהומים, יחד עם שימוש זהירות רבה, כדי לא לגרום נזק מאטר הדורה במהלך הניתוח, תגרום מכוסה זכוכית החלונות מוצלחת וארוכת טווח גולגולתי.

Watch this Scientific Journal Video about A Craniotomy Surgery Procedure for Chronic Brain Imaging at JoVE.com

A Craniotomy Surgery Procedure for Chronic Brain Imaging

זה וידאו פרוטוקול להדגים כיצד השתל מכוסה זכוכית חלון גולגולתי במכרסמים. תכשירים אלה יכולים לשמש כרונית in vivo שני הפוטונים הדמיה של הניאוקורטקס על סולמות זמן של חודשים. זה יכול לשמש גם עבור סוגים אחרים של הדמיה, כולל הדמיה אופטית אות מהותי.

כירורגיה Craniotomy נוהל הדמיה מוחית כרונית

Imaging techniques are becoming increasingly important in the study brain function. Among them, two-photon laser scanning microscopy has emerged as an ...

a-craniotomy-surgery-procedure-for-chronic-brain-imaging

Research

JoVE Journal

Biology

63.8K Views.  University of California, Los Angeles. זה וידאו פרוטוקול להדגים כיצד השתל מכוסה זכוכית חלון גולגולתי במכרסמים. תכשירים אלה יכולים לשמש כרונית in vivo שני הפוטונים הדמיה של הניאוקורטקס על סולמות זמן של חודשים. זה יכול לשמש גם עבור סוגים אחרים של הדמיה, כולל הדמיה אופטית אות מהותי.

Video: A Craniotomy Surgery Procedure for Chronic Brain Imaging

Application of Light-cured Dental Adhesive Resin for Mounting Electrodes or Microdialysis Probes in Chronic Experiments

In chronic recording experiments, self-curing dental acrylic resins have been used as a mounting base of electrodes or microdialysis-probes. Since these acrylics do not bond to the bone, screws have been used as anchors. However, in small experimental animals like finches or mouse, their craniums are very fragile and can not successfully hold the anchors. In this report, we propose a new application of light-curing dental resins for mounting base of electrodes or microdialysis probes in chronic experiments. This material allows direct bonding to the cranium. Therefore, anchor screws are not required and surgical field can be reduced considerably. Past experiences show that the bonding effect maintains more than 2 months. Conventional resin's window of time when the materials are pliable and workable is a few minutes. However, the window of working time for these dental adhesives is significantly wider and adjustable.

In this report, we propose a new application of light-curing dental resins for mounting base of electrodes or microdialysis probes in chronic experiments. This material allows direct bonding to the cranium.

יישום של שרף אור נרפא, דבק שיניים אלקטרודות הרכבה או בדיקות Microdialysis בניסויים כרונית

In chronic recording experiments, self-curing dental acrylic resins have been used as a mounting base of electrodes or microdialysis-probes. Since these ...

Micro-drive Array for Chronic in vivo Recording: Drive Fabrication

Chronic recording of large populations of neurons is a valuable technique for studying the function of neuronal circuits in awake behaving rats. Lightweight recording devices carrying a high density array of tetrodes allow for the simultaneous monitoring of the activity of tens to hundreds of individual neurons. Here we describe a protocol for the fabrication of a micro-drive array with twenty one independently movable micro-drives. This device has been used successfully to record from hippocampal and cortical neurons in our lab. We show how to prepare a custom designed, 3-D printed plastic base that will hold the micro-drives. We demonstrate how to construct the individual micro-drives and how to assemble the complete micro-drive array. Further preparation of the drive array for surgical implantation, such as the fabrication of tetrodes, loading of tetrodes into the drive array and gold-plating, is covered in a subsequent video article.

Micro-drive Array for Chronic in vivo Recording: Drive Fabrication

In this protocol we demonstrate how to fabricate a micro-drive array for chronic electrophysiological recordings in rats.

Micro-Drive Array עבור כרונית In vivo</emהקלטה>: כונן ייצור

Chronic recording of large populations of neurons is a valuable technique for studying the function of neuronal circuits in awake behaving rats. ...

Micro-drive Array for Chronic in vivo Recording: Tetrode Assembly

The tetrode, a bundle of four electrodes, has proven to be a valuable tool for the simultaneous recording of multiple neurons in-vivo.  The differential amplitude of action potential signatures over the channels of a tetrode allows for the isolation of single-unit activity from multi-unit signals.  The ability to precisely control the stereotaxic location and depth of the tetrode is critical for studying coordinated neural activity across brain regions.  In combination with a micro-drive array, it is possible to achieve precise placement and stable control of many tetrodes over the course of days to weeks.  In this protocol, we demonstrate how to fabricate and condition tetrodes using basic tools and materials, install the tetrodes into a multi-drive tetrode array for chronic in-vivo recording in the rat, make ground wire connections to the micro-drive array, and attach a protective cone onto the micro-drive array in order to protect the tetrodes from physical contact with the environment.



Micro-drive Array for Chronic in vivo Recording: Tetrode Assembly

In this protocol we demonstrate how to fabricate and condition tetrodes for use with a micro-drive array, which was designed for chronic electrophysiological recordings in rats. In addition, we illustrate the final stages of micro-drive array construction, which includes installing ground wires and a protective cone.

Micro-Drive Array עבור כרונית In vivo</emהקלטה>: Tetrode האסיפה

The tetrode, a bundle of four electrodes, has proven to be a valuable tool for the simultaneous recording of multiple neurons in-vivo.  The differential ...

Murine Renal Transplantation Procedure

Renal orthotopic transplantation in mice is a technically challenging procedure. Although the first kidney transplants in mice were performed by Russell et al over 30 years ago (1) and refined by Zhang et al years later (2), few people in the world have mastered this procedure. In our laboratory we have successfully performed 1200 orthotopic kidney transplantations with &gt; 90% survival rate. The key points for success include stringent control of reperfusion injury, bleeding and thrombosis, both during the procedure and post-transplantation, and use of 10-0 instead of 11-0 suture for anastomoses.
Post-operative care and treatment of the recipient is extremely important to transplant success and evaluation. All renal graft recipients receive antibiotics in the form of an injection of penicillin immediately post-transplant and sulfatrim in the drinking water continually. Overall animal health is evaluated daily and whole blood creatinine analyses are performed routinely with a portable I-STAT machine to assess graft function.

Renal transplantation in mice is a technically challenging procedure that requires careful post-operative care and treatment for success.

Murine נוהל השתלת כליה

Renal orthotopic transplantation in mice is a technically challenging procedure. Although the first kidney transplants in mice were performed by Russell ...

Live Imaging of the Zebrafish Embryonic Brain by Confocal Microscopy

In this video, we demonstrate the method our lab has developed to analyze the cell shape changes and rearrangements required to bend and fold the developing zebrafish brain (Gutzman et al, 2008). Such analysis affords a new understanding of the underlying cell biology required for development of the 3D structure of the vertebrate brain, and significantly increases our ability to study neural tube morphogenesis. The embryonic zebrafish brain is shaped beginning at 18 hours post fertilization (hpf) as the ventricles within the neuroepithelium inflate. By 24 hpf, the initial steps of neural tube morphogenesis are complete. Using the method described here, embryos at the one cell stage are injected with mRNA encoding membrane-targeted green fluorescent protein (memGFP). After injection and incubation, the embryo, now between 18 and 24 hpf, is mounted, inverted, in agarose and imaged by confocal microscopy. Notably, the zebrafish embryo is transparent making it an ideal system for fluorescent imaging. While our analyses have focused on the midbrain-hindbrain boundary and the hindbrain, this method could be extended for analysis of any region in the zebrafish to a depth of 
80-100 &mu;m.

In this video, we demonstrate a method by which to analyze the developing vertebrate brain in live zebrafish embryos at single cell resolution by confocal microscopy. This includes the method by which we inject the single-cell zebrafish embryo and subsequently mount and image the developing brain.

חיים הדמיה של המוח על ידי עוברי דג הזברה מיקרוסקופיה confocal

In this video, we demonstrate the method our lab has developed to analyze the cell shape changes and rearrangements required to bend and fold the ...

Making MR Imaging Child's Play - Pediatric Neuroimaging Protocol, Guidelines and Procedure

Within the last decade there has been an increase in the use of structural and functional magnetic resonance imaging (fMRI) to investigate the neural basis of human perception, cognition and behavior 1, 2. Moreover, this non-invasive imaging method has grown into a tool for clinicians and researchers to explore typical and atypical brain development. Although advances in neuroimaging tools and techniques are apparent, (f)MRI in young pediatric populations remains relatively infrequent 2. Practical as well as technical challenges when imaging children present clinicians and research teams with a unique set of problems 3, 2. To name just a few, the child participants are challenged by a need for motivation, alertness and cooperation. Anxiety may be an additional factor to be addressed. Researchers or clinicians need to consider time constraints, movement restriction, scanner background noise and unfamiliarity with the MR scanner environment2,4-10. A progressive use of functional and structural neuroimaging in younger age groups, however, could further add to our understanding of brain development. As an example, several research groups are currently working towards early detection of developmental disorders, potentially even before children present associated behavioral characteristics e.g.11. Various strategies and techniques have been reported as a means to ensure comfort and cooperation of young children during neuroimaging sessions. Play therapy 12, behavioral approaches 13, 14,15, 16-18 and simulation 19, the use of mock scanner areas 20,21, basic relaxation 22 and a combination of these techniques 23 have all been shown to improve the participant's compliance and thus MRI data quality. Even more importantly, these strategies have proven to increase the comfort of families and children involved 12. One of the main advances of such techniques for the clinical practice is the possibility of avoiding sedation or general anesthesia (GA) as a way to manage children's compliance during MR imaging sessions 19,20. In the current video report, we present a pediatric neuroimaging protocol with guidelines and procedures that have proven to be successful to date in young children.

Despite an increase in the use of structural and functional magnetic resonance imaging (fMRI) in humans, the study of young pediatric populations remains a challenge. We present a hands-on, step-by-step video protocol including guidelines for clinicians and researchers intending to perform (f)MRI in young children.

ביצוע לשחק MR הילד הדמיה - פרוטוקול הדמייה ילדים, הנחיות נוהל

Within the last decade there has been an increase in the use of structural and functional magnetic resonance imaging (fMRI) to investigate the neural ...

Bioluminescence Imaging of Heme Oxygenase-1 Upregulation in the Gua Sha Procedure

Gua Sha is a traditional Chinese folk therapy that employs skin scraping to cause subcutaneous microvascular blood extravasation and bruises. The protocol for bioluminescent optical imaging of HO-1-luciferase transgenic mice reported in this manuscript provides a rapid in vivo assay of the upregulation of the heme oxygenase-1 (HO-1) gene expression in response to the Gua Sha procedure. HO-1 has long been known to provide cytoprotection against oxidative stress. The upregulation of HO-1, assessed by the bioluminescence output, is thought to represent an antioxidative response to circulating hemoglobin products released by Gua Sha. Gua Sha was administered by repeated strokes of a smooth spoon edge over lubricated skin on the back or other targeted body part of the transgenic mouse until petechiae (splinter hemorrhages) or ecchymosis (bruises) indicative of extravasation of blood from subcutaneous capillaries was observed. After Gua Sha, bioluminescence imaging sessions were carried out daily for several days to follow the dynamics of HO-1 expression in multiple internal organs.

Gua Sha, traditional Chinese therapeutic skin scraping, causes subcutaneous microvascular blood extravasation. We report a protocol of bioluminescence imaging of HO-1-luciferase transgenic mice to demonstrate that Gua Sha upregulates heme oxygenase-1 (HO-1) in multiple organs.

הדמיה של פליטת אור upregulation הרן-1 Oxygenase בנוהל גואה שא

Gua Sha is a traditional Chinese folk therapy that employs skin scraping to cause subcutaneous microvascular blood extravasation and bruises. The protocol ...

Mapping the After-effects of Theta Burst Stimulation on the Human Auditory Cortex with Functional Imaging

Auditory cortex pertains to the processing of sound, which is at the basis of speech or music-related processing1. However, despite considerable recent progress, the functional properties and lateralization of the human auditory cortex are far from being fully understood. Transcranial Magnetic Stimulation (TMS) is a non-invasive technique that can transiently or lastingly modulate cortical excitability via the application of localized magnetic field pulses, and represents a unique method of exploring plasticity and connectivity. It has only recently begun to be applied to understand auditory cortical function 2. 
An important issue in using TMS is that the physiological consequences of the stimulation are difficult to establish. Although many TMS studies make the implicit assumption that the area targeted by the coil is the area affected, this need not be the case, particularly for complex cognitive functions which depend on interactions across many brain regions 3. One solution to this problem is to combine TMS with functional Magnetic resonance imaging (fMRI). The idea here is that fMRI will provide an index of changes in brain activity associated with TMS. Thus, fMRI would give an independent means of assessing which areas are affected by TMS and how they are modulated 4. In addition, fMRI allows the assessment of functional connectivity, which represents a measure of the temporal coupling between distant regions. It can thus be useful not only to measure the net activity modulation induced by TMS in given locations, but also the degree to which the network properties are affected by TMS, via any observed changes in functional connectivity.
Different approaches exist to combine TMS and functional imaging according to the temporal order of the methods. Functional MRI can be applied before, during, after, or both before and after TMS. Recently, some studies interleaved TMS and fMRI in order to provide online mapping of the functional changes induced by TMS 5-7. However, this online combination has many technical problems, including the static artifacts resulting from the presence of the TMS coil in the scanner room, or the effects of TMS pulses on the process of MR image formation. But more importantly, the loud acoustic noise induced by TMS (increased compared with standard use because of the resonance of the scanner bore) and the increased TMS coil vibrations (caused by the strong mechanical forces due to the static magnetic field of the MR scanner) constitute a crucial problem when studying auditory processing. 
This is one reason why fMRI was carried out before and after TMS in the present study. Similar approaches have been used to target the motor cortex 8,9, premotor cortex 10, primary somatosensory cortex 11,12 and language-related areas 13, but so far no combined TMS-fMRI study has investigated the auditory cortex. The purpose of this article is to provide details concerning the protocol and considerations necessary to successfully combine these two neuroscientific tools to investigate auditory processing. 
Previously we showed that repetitive TMS (rTMS) at high and low frequencies (resp. 10 Hz and 1 Hz) applied over the auditory cortex modulated response time (RT) in a melody discrimination task 2. We also showed that RT modulation was correlated with functional connectivity in the auditory network assessed using fMRI: the higher the functional connectivity between left and right auditory cortices during task performance, the higher the facilitatory effect (i.e. decreased RT) observed with rTMS. However those findings were mainly correlational, as fMRI was performed before rTMS. Here, fMRI was carried out before and immediately after TMS to provide direct measures of the functional organization of the auditory cortex, and more specifically of the plastic reorganization of the auditory neural network occurring after the neural intervention provided by TMS. 
Combined fMRI and TMS applied over the auditory cortex should enable a better understanding of brain mechanisms of auditory processing, providing physiological information about functional effects of TMS. This knowledge could be useful for many cognitive neuroscience applications, as well as for optimizing therapeutic applications of TMS, particularly in auditory-related disorders.

Auditory processing is the basis of speech and music-related processing. Transcranial Magnetic Stimulation (TMS) has been used successfully to study cognitive, sensory and motor systems but has rarely been applied to audition. Here we investigated TMS combined with functional Magnetic Resonance Imaging to understand the functional organization of auditory cortex.

מיפוי לאחר הלוואי של תטא Burst גירוי בקורטקס השמיעתי האנושי בהדמיה תפקודית

Auditory cortex pertains to the processing of sound, which is at the basis of speech or music-related processing1. However, despite considerable recent ...

Preparation of Pancreatic Acinar Cells for the Purpose of Calcium Imaging, Cell Injury Measurements, and Adenoviral Infection

The pancreatic acinar cell is the main parenchymal cell of the exocrine pancreas and plays a primary role in the secretion of pancreatic enzymes into the pancreatic duct. It is also the site for the initiation of pancreatitis. Here we describe how acinar cells are isolated from whole pancreas tissue and intracellular calcium signals are measured. In addition, we describe the techniques of transfecting these cells with adenoviral constructs, and subsequently measuring the leakage of lactate dehydrogenase, a marker of cell injury, during conditions that induce acinar cell injury in vitro. These techniques provide a powerful tool to characterize acinar cell physiology and pathology.

We describe a reproducible method of preparing mouse pancreatic acinar cells from a mouse for the purpose of examining acinar cell calcium signals and cellular injury with physiologically and pathologically relevant stimuli. A method for adenoviral infection of these cells is also provided.

הכנת תאי לבלב acinar לצורך סידן ההדמיה, פגיעה בתא המדידות, וזיהום adenoviral

The pancreatic acinar  cell is the main parenchymal cell of the exocrine pancreas and plays a primary  role in the secretion of pancreatic enzymes into ...

Isolation of Human Hepatocytes by a Two-step Collagenase Perfusion Procedure

The liver, an organ with an exceptional regeneration capacity, carries out a wide range of functions, such as detoxification, metabolism and homeostasis. As such, hepatocytes are an important model for a large variety of research questions. In particular, the use of human hepatocytes is especially important in the fields of pharmacokinetics, toxicology, liver regeneration and translational research. Thus, this method presents a modified version of a two-step collagenase perfusion procedure to isolate hepatocytes as described by Seglen 1.
Previously, hepatocytes have been isolated by mechanical methods. However, enzymatic methods have been shown to be superior as hepatocytes retain their structural integrity and function after isolation. This method presented here adapts the method designed previously for rat livers to human liver pieces and results in a large yield of hepatocytes with a viability of 77&plusmn;10%. The main difference in this procedure is the process of cannulization of the blood vessels. Further, the method described here can also be applied to livers from other species with comparable liver or blood vessel sizes.

A modified two-step collagenase perfusion procedure for isolation of human hepatocytes is described. This method can also be applied to other mammalian livers. The isolated hepatocytes are available in high yield and viability, making them a suitable model for scientific research in areas such as liver regeneration, pharmacokinetics and toxicology.

בידוד של hepatocytes האנושי על ידי שני שלבים Collagenase זלוף נוהל

The liver, an organ with an exceptional regeneration capacity, carries out a wide range of functions, such as detoxification, metabolism and homeostasis. ...

כירורגיה Craniotomy נוהל הדמיה מוחית כרונית

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